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Development of Low-Immunogenic AFPs for Cell Cryopreservation by Hydrophilic Modification
Feng Shen1,2, Xiaoying Jiang1,2, Andrew B Zhu3
1Department of Biochemical Engineering, School of Chemical Engineering and Technology, Tianjin University, Tianjin 300350, China.
Langmuir : the ACS Journal of Surfaces and Colloids
|July 22, 2025
Summary
Researchers developed low-immunogenic antifreeze proteins (AFPs) for cryopreservation by hydrophilic modification. These modified AFPs show reduced immune response and high cell survival rates, paving the way for advanced cell and tissue preservation.
Area of Science:
- Biochemistry
- Cryobiology
- Immunology
Background:
- Antifreeze proteins (AFPs) are crucial for cryopreservation.
- Their clinical application is hindered by immunogenicity.
- Hydrophilic modification offers a strategy to mitigate immune responses.
Purpose of the Study:
- To develop low-immunogenic antifreeze proteins (AFPs) using hydrophilic modification.
- To evaluate the immunogenicity and biocompatibility of modified AFPs.
- To assess the efficacy of modified AFPs in cell cryopreservation.
Main Methods:
- Design and synthesis of hydrophilic polypeptides (KE and GES).
- Conjugation of polypeptides to natural Marinomonas primoryensis AFPs (MpAFPs) to create AKE and AGES.
- In vivo immunogenicity testing and biocompatibility assessment in a mouse model.
- Cryopreservation of NIH/3T3 cells using modified AFPs.
Main Results:
- AKE and AGES demonstrated significantly reduced immunogenicity (48% and 42% lower, respectively) compared to native MpAFP.
- Biocompatibility tests revealed negligible adverse effects on liver, heart, and kidney functions.
- Successful cryopreservation of NIH/3T3 cells with high survival rates (96% for AKE, 87% for AGES).
Conclusions:
- Hydrophilic modification effectively reduces AFP immunogenicity.
- Modified AFPs (AKE and AGES) are biocompatible and efficient for cell cryopreservation.
- This work establishes a foundation for next-generation low-immunogenic AFPs in regenerative medicine and tissue banking.
